The COVID-19 diagnosis was confirmed if patients tested positive for SARS-CoV-2 by PCR of nasal or pharyngeal swab specimens (majority of tests by Roche, Basel, Switzerland; 99.1% sensitivity and 99.7% specificity; other tests used with lower frequency included: DiaSorin: 500 copies/mL; Thermo Fisher: 10 genomic copy equivalents/reaction; Seegene: 1250 copies/mL; Hologic: TCID50/mL: 1 × 10–2) or retrospectively by IgG detection tests [DiaSorin: sensitivity 97.6% (≥15 days after diagnosis), specificity 99.3%; Diazyme: sensitivity 91.2%, specificity 97.3%]. Only patients who had a record of a positive test result were included in the analysis. The PCR assays were authorised by the US Food and Drug Administration (FDA) without clinical sensitivity/specificity data owing to the urgent nature of the pandemic. Only one positive test was necessary for the patient to be included in the retrospective analysis.
Tcid50
Manufactured by Thermo Fisher Scientific
Sourced in Switzerland
The TCID50 is a laboratory assay used to determine the titer or concentration of a virus or other infectious agent. It measures the dilution at which 50% of cell cultures or test organisms become infected by the agent. The TCID50 provides a quantitative assessment of the infectivity of a viral preparation.
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Lab products found in correlation
2 protocols using tcid50
1
SARS-CoV-2 Diagnostic Testing Protocol
2
Purification of Infectious EV71 Particles
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It is well known that two forms of EV71 viral particles, full and empty particles, existed during propagation in cells [3] (link), [26] (link). Based on historical poliovirus studies, the full particles are infectious and immunogenic but the empty particles are not [27] (link). Therefore, we purified EV71 infectious (full) particle of the reference viruses for rabbit immunizations. The EV71 culture supernatant was concentrated 10-fold with a Amicon 100K centrifugal filter (Millipore). The crude virus concentrate was loaded onto a 15–65% continuous sucrose gradient and centrifuged at 28000 rpm for 4 hr. Fractions (2 mL per fraction) were collected and the viral titer and protein concentration of each fraction were determined by TCID50 and BCA assays (Thermo Scientific), respectively. Fractions with high infectious virus titers in 32–38% sucrose concentration were merged and concentrated by diafiltration using Amicon 100K centrifugal filter and centrifugation at 3500 g. The purified EV71 viruses were further verified by using Western blot and electron microscopy analysis.
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